Affinage

RPE65

Retinoid isomerohydrolase · UniProt Q16518

Length
533 aa
Mass
60.9 kDa
Annotated
2026-06-10
100 papers in source corpus 32 papers cited in narrative 32 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 5/6 claims corpus-supported (83%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

RPE65 is the retinal pigment epithelium-specific retinoid isomerohydrolase that performs the rate-limiting step of the visual cycle, converting all-trans-retinyl esters into 11-cis-retinol for regeneration of the visual chromophore (PMID:16116091, PMID:16096063, PMID:16026160). Genetic ablation in mice abolishes 11-cis-retinoid production and rhodopsin, accumulates all-trans-retinyl esters in the RPE, and eliminates rod ERG responses, establishing RPE65 as indispensable for chromophore supply (PMID:9843205); epistasis in cone-only and rod-null backgrounds shows it is equally required for cone vision, with foveal cones being especially dependent on robust RPE65 activity (PMID:17251447, PMID:17848510). Catalysis requires LRAT to generate the all-trans-retinyl ester substrate, and RPE65 functions within an RPE microsomal multiprotein complex together with RDH5 and RGR (PMID:16116091, PMID:20100834). Although it lacks transmembrane segments, RPE65 associates peripherally with ER/microsomal membranes through hydrophobic and phospholipid interactions and through S-palmitoylation at C112 and C146, with membrane integrity being essential for isomerase activity (PMID:9328280, PMID:20100834, PMID:30914787). RPE65 activity is quantitatively rate-limiting and tunable: it is negatively regulated by FATP4 through competition for the retinyl ester substrate, with very-long-chain acyl-CoAs inhibiting and palmitoyl-CoA promoting activity (PMID:23407971, PMID:16026160). RPE65 loss-of-function and instability cause inherited retinal dystrophy: most disease-associated missense mutations abolish isomerase activity or destabilize the protein, and non-active-site misfolding mutants are cleared by PSMD13-mediated ubiquitin-proteasome degradation, whereas the dominant D477G mutation delays chromophore regeneration through a toxic/dominant-negative mechanism involving abnormal complex formation (PMID:16754667, PMID:24849605, PMID:29659842, PMID:19431183).

Mechanistic history

Synthesis pass · year-by-year structured walk · 12 steps
  1. 1993 High

    Established RPE65 as an RPE-specific protein and raised the question of how a soluble-looking protein associates with membranes, setting up the membrane-association and post-translational regulation problems.

    Evidence cDNA cloning from bovine RPE, Northern/Western blot, peptide microsequencing

    PMID:8340400

    Open questions at the time
    • Did not assign a catalytic function
    • Nature of the higher-mass membrane modification undefined
    • Mechanism of post-transcriptional regulation in cultured RPE not identified
  2. 1998 High

    Defined RPE65's physiological role by showing its loss blocks 11-cis-retinoid production and rhodopsin formation, identifying it as a required node in the retinal visual cycle.

    Evidence Rpe65 knockout mice with ERG, HPLC retinoid analysis, immunohistochemistry; corroborated by canine frameshift model

    PMID:9808841 PMID:9843205

    Open questions at the time
    • Did not establish the biochemical reaction RPE65 catalyzes
    • Direct enzyme activity not demonstrated
    • Role in cones not yet tested
  3. 1997 Medium

    Showed RPE65 is a peripheral membrane protein associating with microsomes via phospholipid/hydrophobic interactions, answering how a protein lacking transmembrane domains localizes to membranes.

    Evidence Liposome co-sedimentation, IAM chromatography, resonance energy transfer

    PMID:9328280

    Open questions at the time
    • Specific residues/modifications mediating association not mapped
    • Single-lab biochemistry
    • Functional consequence of membrane association untested here
  4. 2005 High

    Resolved RPE65's molecular activity by directly demonstrating isomerohydrolase catalysis of all-trans-retinyl ester to 11-cis-retinol, and showed it depends on LRAT for substrate and is quantitatively rate-limiting in vivo.

    Evidence Recombinant expression enzymatic assays in mammalian/insect cells with HPLC, unbiased cDNA screen, mutagenesis of LCA residues, in vivo Michaelis-Menten kinetics; rd12 nonsense model

    PMID:15765048 PMID:16026160 PMID:16096063 PMID:16116091

    Open questions at the time
    • Catalytic mechanism/cofactor requirements not fully defined
    • Structure not solved
    • Relationship to membrane association mechanistically incomplete
  5. 2007 High

    Demonstrated RPE65 is required for the cone as well as rod visual cycle, addressing whether cones use an RPE65-independent chromophore source.

    Evidence Rpe65 ablation in Nrl-/- and Rho-/- mice with ERG and HPLC; gene-delivery rescue preventing cone degeneration; cone expression and central RPE enrichment data

    PMID:11980880 PMID:16505056 PMID:17251447 PMID:17848510 PMID:22171060

    Open questions at the time
    • Whether cone-intrinsic RPE65 contributes vs RPE-derived chromophore not resolved
    • Cone-specific alternative pathway in some species (zebrafish) unexplained
    • Quantitative cone vs rod chromophore demand undefined
  6. 2018 Medium

    Tested whether RPE65 expressed within cones is functionally relevant, refining where chromophore-regenerating activity matters.

    Evidence Cone-specific human RPE65 transgenic mice on RPE65-deficient background; ERG, single-cell electrophysiology; zebrafish knockdown with retinylamine epistasis

    PMID:17868371 PMID:30242264

    Open questions at the time
    • Negative result in mouse may not generalize across species
    • Identity of the RPE65-independent cone pathway unknown
    • Buffering vs catalytic role of cone RPE65 unresolved
  7. 2006 High

    Connected disease mutations to mechanism by showing inactivating mutations abolish isomerase activity and destabilize protein while shifting it off membranes, defining loss-of-function as the dominant disease route.

    Evidence In vitro isomerase assays, subretinal rescue in Rpe65-/- mice, cycloheximide-chase stability, fractionation; systematic functional testing of variant series; homology modeling

    PMID:16754667 PMID:18599565 PMID:18722466 PMID:19431183

    Open questions at the time
    • Per-residue catalytic contributions not fully mapped without a crystal structure
    • Relationship between stability loss and membrane displacement mechanistically incomplete
    • Residual-activity thresholds for phenotype only correlative
  8. 2014 High

    Identified the quality-control pathway disposing of misfolded RPE65, showing PSMD13 mediates ubiquitin-proteasome degradation of misfolding mutants and that chemical chaperones rescue non-active-site but not active-site mutants.

    Evidence Co-IP, ubiquitination assay, proteasome inhibition, isomerase activity, low-temperature and 4-PBA/glycerol rescue; rd12 translational-suppression analysis

    PMID:24644049 PMID:24849605

    Open questions at the time
    • Whether PSMD13 acts via canonical 26S targeting or a dedicated recognition step unclear
    • In vivo relevance of this pathway not tested
    • E3 ligase for RPE65 not identified
  9. 2013 High

    Revealed metabolic regulation of RPE65 by showing FATP4 negatively modulates isomerase activity through substrate competition and acyl-CoA chain-length effects, linking chromophore regeneration speed to lipid metabolism.

    Evidence Unbiased bovine RPE cDNA screen, FATP4 knockout mice, isomerase and ERG recovery kinetics, substrate competition assays

    PMID:23407971

    Open questions at the time
    • Physiological conditions that engage FATP4 regulation undefined
    • Whether ELOVL1 or other lipid enzymes participate not tested here
    • Stoichiometry of competition at the membrane unknown
  10. 2019 High

    Defined S-palmitoylation as a regulated determinant of RPE65 membrane association, mapping modified cysteines and linking palmitoylation dynamics to LRAT substrate availability.

    Evidence Acyl-RAC palmitoylation assay, palmitoylation inhibitors, C112/C146 mutagenesis, membrane fractionation, LRAT co-expression

    PMID:30914787

    Open questions at the time
    • Palmitoyltransferase responsible not identified
    • Functional consequence on catalytic rate vs localization not fully separated
    • In vivo significance of dynamic palmitoylation untested
  11. 2019 High

    Mechanistically dissected the dominant D477G mutation, showing it acts through a toxic gain-of-function/dominant-negative route (aggregation-prone surface, abnormal complex formation, delayed regeneration) plus a human splicing defect, distinguishing it from recessive loss-of-function.

    Evidence D477G knock-in mice with HPLC/ERG/ubiquitination, cell-line activity assay, structural analysis, heterozygote dominant-negative kinetics, Exontrap splicing assay

    PMID:28041994 PMID:29659842 PMID:30628748

    Open questions at the time
    • Relative contributions of splicing defect vs protein-level toxicity in humans unresolved
    • Composition of the abnormal complex not defined
    • Whether the same mechanism operates in human RPE not directly shown
  12. 2018 Medium

    Established pharmacological tractability of the substrate-binding site by developing a competitive non-retinoid inhibitor that slows chromophore regeneration and protects against light damage.

    Evidence In vitro competitive inhibition kinetics (CU239), in vivo HPLC and ERG light-damage model in mice

    PMID:29684583

    Open questions at the time
    • Binding mode not structurally resolved
    • Therapeutic window and selectivity in humans untested
    • Long-term consequences of chronic inhibition unknown

Open questions

Synthesis pass · forward-looking unresolved questions
  • How RPE65, LRAT, RDH5, RGR and lipid regulators are spatially organized into a functional membrane-bound visual-cycle machine, and the high-resolution catalytic mechanism of isomerization, remain incompletely defined.
  • No experimentally determined high-resolution structure of human RPE65 in the timeline
  • Architecture and stoichiometry of the RDH5/RGR complex undefined
  • Identity of palmitoyltransferase and ubiquitin ligase acting on RPE65 unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016853 isomerase activity 5 GO:0008289 lipid binding 2 GO:0016787 hydrolase activity 2
Localization
GO:0005783 endoplasmic reticulum 3 GO:0005829 cytosol 2 GO:0005886 plasma membrane 2
Pathway
R-HSA-9709957 Sensory Perception 3 R-HSA-1430728 Metabolism 2
Partners
FATP4LRATPSMD13RDH5RGR
Complex memberships
RPE65-RDH5-RGR RPE microsomal isomerization complex

Evidence

Reading pass · 32 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1998 Rpe65-deficient mice lack rhodopsin (but retain opsin apoprotein), have abolished rod ERG responses, accumulate all-trans-retinyl esters in the RPE, and lack 11-cis-retinyl esters, establishing that RPE65 is necessary for production of 11-cis-vitamin A in the retinal visual cycle. Rpe65 knockout mouse model; electroretinography; retinoid biochemical analysis by HPLC; immunohistochemistry Nature genetics High 9843205
2005 Recombinant RPE65, when expressed in QBI-293A and COS-1 cells, catalyzes the conversion of all-trans-retinyl ester to 11-cis-retinol (isomerohydrolase activity) at a rate of ~2.9 pmol/min per mg; activity requires co-expression of lecithin retinol acyltransferase (LRAT) to provide substrate, establishing RPE65 as the visual cycle isomerohydrolase. Cell-based enzymatic assay (recombinant expression in QBI-293A and COS-1 cells); HPLC retinoid analysis; dose-response correlation with RPE65 expression levels Proceedings of the National Academy of Sciences of the United States of America High 16116091
2005 An unbiased cDNA expression screen identified Rpe65 as the retinoid isomerase converting all-trans-retinyl ester to 11-cis-retinol; Leber-associated missense mutations C330Y and Y368H abolished isomerase activity, confirming catalytic identity. Unbiased cDNA expression screen; catalytic activity assay in mammalian and insect cells; site-directed mutagenesis of disease-associated residues Cell High 16096063
1993 RPE65 was molecularly cloned from bovine RPE; the 533-amino-acid protein lacks transmembrane segments, is RPE-specific by Northern blot, and the microsomal membrane-associated form has a higher apparent molecular mass than the soluble form, suggesting post-translational modification; cultured RPE cells contain RPE65 mRNA but no immunodetectable protein, indicating post-transcriptional regulation. cDNA library screening; RACE; heterologous expression in E. coli; Western blot; Northern blot; peptide microsequencing The Journal of biological chemistry High 8340400
2001 RPE65 exists in two forms in bovine RPE: a cytosolic form with mass close to calculated (~61 kDa) and a microsomal membrane-associated form with higher mass (~62 kDa), indicating the membrane-associated form carries post-translational modifications; recombinant human RPE65 expressed in Sf9 cells likewise shows membrane-associated and non-membrane forms with differing masses. Baculovirus/Sf9 expression; affinity chromatography purification; MALDI mass spectrometry; subcellular fractionation; Western blot; immunocytochemistry Investigative ophthalmology & visual science Medium 11381042
1997 RPE65 lacks transmembrane domains yet associates with RPE microsomal membranes through hydrophobic/phospholipid interactions; RPE65 co-sediments with phosphatidylcholine and other phospholipid liposomes in a Ca2+-independent manner and binds PC-IAM matrix, demonstrating peripheral membrane association via phospholipid binding. Phospholipid liposome co-sedimentation assay; IAM chromatography; resonance energy transfer; spectrophotometric aggregation assay Archives of biochemistry and biophysics Medium 9328280
2010 Hydrophobic interactions are the dominant forces promoting RPE65 association with native RPE microsomal membranes; phospholipid membrane structural integrity is critical for RPE65 isomerization activity (phospholipase A2 treatment inhibits 11-cis-retinol production in correlation with lipid hydrolysis kinetics); RPE65 operates in a multiprotein complex with retinol dehydrogenase 5 (RDH5) and retinal G protein-coupled receptor (RGR) in RPE microsomes. Membrane extraction and phase separation; phospholipase treatment with isomerase activity assay; co-immunoprecipitation from RPE microsomes The Journal of biological chemistry Medium 20100834
2006 Disease-associated RPE65 mutations R91W and Y368H both abolish isomerohydrolase activity in vitro and in vivo (subretinal injection into Rpe65-/- mice); these mutants show decreased protein stability (half-lives <2 h and <6 h vs. >10 h for WT) and shift from membrane to cytosolic fractions, indicating the mutations disrupt membrane association and reduce stability; palmitoylation is retained in both mutants. In vitro isomerohydrolase assay; subretinal injection rescue experiment in Rpe65-/- mice; protein stability/cycloheximide chase; subcellular fractionation; Western blot; palmitoylation assay The Journal of biological chemistry High 16754667
2005 Adenovirus-mediated delivery of RPE65 to Rpe65-/- mice restores isomerohydrolase activity in eyecup homogenates to wild-type levels, restores normal retinoid profile (including 11-cis-retinal), and prevents early cone degeneration, confirming that RPE65 isomerohydrolase activity is required for cone photoreceptor survival. Subretinal adenovirus injection; in vitro isomerohydrolase activity assay; HPLC retinoid profiling; immunohistochemistry; cone counting on flatmounted retina; RT-PCR Investigative ophthalmology & visual science High 16505056
2007 RPE65 is essential for production of 11-cis-retinal for cone photoreceptors: ablation of RPE65 in Nrl-/- (cone-only) and Rho-/- mice eliminates 11-cis-retinal and causes a ~1000-fold drop in retinal sensitivity, demonstrating RPE65 is indispensable for cone as well as rod visual cycle function. Genetic crosses (Rpe65-/- × Nrl-/- and Rpe65-/- × Rho-/-); electroretinography; HPLC retinoid profiling; Western blot; immunohistochemistry Investigative ophthalmology & visual science High 17251447
2002 RPE65 protein is expressed in mammalian cone photoreceptors (mouse, rabbit, cow, Xenopus) but not in rods, as determined by immunohistochemistry on flatmounted and sectioned retinas; double-labeling with PNA confirmed cone identity, and RPE65 knockout confirmed antibody specificity. Immunohistochemistry on flatmounted and sectioned retinas; double-labeling with PNA; Western blot; RPE65 knockout control Investigative ophthalmology & visual science Medium 11980880
1998 RPE65 mRNA is expressed in salamander single cone cells (but not rods), as detected by RT-PCR on isolated cells, suggesting RPE65 contributes to the unique retinoid processing pathway in cones. RT-PCR on individually isolated photoreceptor cells; DNA sequencing confirmation Biochimica et biophysica acta Low 9838153
2011 RPE65 is present in human green/red cones but absent from blue cones; in the 661W cone cell line, RPE65 mediates ester hydrolysis for photopigment synthesis in vitro. Immunohistochemistry on human retinal sections; in vitro ester hydrolysis assay in 661W cone cell line The Journal of neuroscience : the official journal of the Society for Neuroscience Medium 22171060
2007 RPE65 retinoid isomerase activity is ~4-fold higher in central versus peripheral macaque RPE by direct enzymatic assay, and RPE65 concentrates in central retina by immunoblotting; early cone photoreceptor loss in RPE65-LCA patients indicates robust RPE65-based chromophore production is important for foveal cone survival. Retinoid isomerase activity assay on macaque RPE fractions; immunoblotting; immunocytochemistry; in vivo retinal imaging of human patients Proceedings of the National Academy of Sciences of the United States of America Medium 17848510
2008 The hypomorphic RPE65 mutation P25L reduces isomerohydrolase activity to 7.7% of wild-type in transfected 293F cells, and the L22P mutation to 13.5%; this residual activity correlates with a milder clinical phenotype (preserved cone function), establishing a dose-response relationship between RPE65 isomerase activity level and retinal functional outcome. Cell-based isomerase activity assay (293F cells); HPLC retinoid quantification; clinical ERG and psychophysics Investigative ophthalmology & visual science Medium 18599565
2008 The novel RPE65 mutation G244V (near the catalytic center) causes protein instability in cultured cells and abolishes RPE65-dependent isomerase activity; homology modeling using the carotenoid oxygenase ACO structure localizes known inactivating mutations around the catalytic region. Cell-based isomerase assay; EGFP bicistronic expression + cell sorting + immunoblotting; homology modeling Biochemistry Medium 18722466
2014 Three disease-associated RPE65 missense mutations (L22P, T101I, L408P) at non-active sites cause misfolding and aggregation via disulfide bonds; PSMD13 (26S proteasome non-ATPase regulatory subunit 13) interacts with and mediates rapid ubiquitination/proteasome-dependent degradation of misfolded (but not properly folded) mutant RPE65 proteins; low temperature, sodium 4-phenylbutyrate, and glycerol can rescue catalytic activity of non-active-site mutants but not active-site mutants. Co-immunoprecipitation; ubiquitination assay; proteasome inhibitor experiments; isomerase activity assay; low-temperature rescue assay; chemical chaperone treatment The Journal of biological chemistry High 24849605
2013 FATP4 (fatty acid transport protein 4) is a negative regulator of RPE65 isomerase activity; lignoceroyl (C24:0)-CoA inhibits 11-cis-retinol synthesis whereas palmitoyl (C16:0)-CoA promotes it; FATP4-deficient RPE shows higher isomerase activity, faster 11-cis-retinaldehyde regeneration, and faster rod light sensitivity recovery; FATP4 competes with RPE65 for its substrate all-trans-retinyl ester. Unbiased bovine RPE cDNA expression screen; FATP4 knockout mice; isomerase activity assay; ERG recovery kinetics; substrate competition assay The Journal of neuroscience : the official journal of the Society for Neuroscience High 23407971
2019 RPE65 undergoes S-palmitoylation (~25% of total protein) at residues C112 and C146; inhibition of palmitoylation abolishes membrane association; palmitoylation-deficient C112 mutants show impaired membrane association; LRAT dynamically regulates RPE65 palmitoylation level (decreased in the presence of all-trans-retinol, the LRAT substrate). Palmitoylation assay (acyl-RAC); 2-bromopalmitate and 2-fluoropalmitate inhibition; site-directed mutagenesis; membrane fractionation; LRAT co-expression experiments Scientific reports High 30914787
2018 The dominant D477G RPE65 mutation generates an aggregation-prone surface; knock-in mice show ubiquitinated and reduced RPE65, retinyl ester accumulation, delayed rhodopsin regeneration kinetics, and diminished ERG responses; a cell line expressing D477G shows normal localization and isomerase activity, suggesting a toxic gain-of-function through abnormal complex formation rather than simple loss-of-function. D477G knock-in mouse model; immunoblot for ubiquitination; HPLC retinoid analysis; ERG; cell line isomerase activity assay; structural analysis of RPE65 chimera and crystal packing Human molecular genetics High 29659842
2016 The D477G dominant mutation acts as a dominant-negative: heterozygous knock-in mice show slower kinetics of 11-cis-retinal regeneration after light exposure and delayed dark adaptation (lower A-wave recovery after photobleaching) compared with both wild-type and RPE65 heterozygous knockout mice, despite comparable steady-state 11-cis-retinal levels. D477G knock-in mouse; ERG photobleach-recovery kinetics; HPLC retinoid analysis The American journal of pathology Medium 28041994
2019 The human RPE65 c.1430A>G (D477G) mutation generates an ectopic splice site causing aberrant splicing of RPE65 mRNA, disrupting protein expression; this splicing defect was confirmed for the human mutant in an in vitro Exontrap assay; homozygous knock-in mice show light-stress-induced retinal degeneration. CRISPR/Cas9 knock-in mouse; Exontrap in vitro splicing assay; ERG; retinal histology; mRNA analysis Human mutation Medium 30628748
2010 RPE65 cleavage occurs under oxidative stress: H2O2 exposure of RPE cells generates a novel 45 kDa truncated fragment (RPE45) via a caspase-mediated, ubiquitination-dependent mechanism; the same cleavage occurs in vivo upon light exposure in mice. Western blot; recombinant caspase cleavage assay; mass spectrometry; in vivo light-exposure experiment International journal of biological macromolecules Low 20510285
2006 The Rpe65 L450M/L450 sequence variant acts as a genetic modifier of retinal degeneration in a transgenic RP mouse model: mice with the Rpe65(450Met) variant retain more rhodopsin and show reduced retinal degeneration compared to mice with the Rpe65(450Leu) variant; phototransduction (not c-Fos) pathway mediates this light-driven degeneration. Transgenic mouse crosses with different Rpe65 variants; ERG; rhodopsin quantification; genetic epistasis (c-Fos KO and phototransduction mutant crosses) The European journal of neuroscience Medium 16519667
2005 In vivo, the initial rate of 11-cis-retinal synthesis is a Michaelis function of RPE65 quantity (Vmax ~18 pmol/min per eye, Km ~1.7 pmol); at sub-Km RPE65 levels, each RPE65 molecule supports ~10 molecules of 11-cis-retinal per minute regardless of the L450/M450 variant, establishing RPE65 as a quantitative rate-limiting factor for rhodopsin regeneration. Quantitative ELISA for RPE65 per eye; in vivo rhodopsin regeneration kinetics measured by ERG after bleach in five genotypes; Michaelis-Menten kinetic fitting Biochemistry Medium 16026160
2018 A novel non-retinoid small molecule inhibitor CU239 selectively inhibits RPE65 isomerase activity (IC50 ~6 μM) via competition with its all-trans-retinyl ester substrate; systemic injection in mice delays chromophore regeneration and partially protects the retina from high-intensity light damage. In vitro isomerase activity assay; competitive inhibition kinetics; in vivo retinoid analysis (HPLC); ERG light-damage model in mice Biochimica et biophysica acta. Molecular basis of disease Medium 29684583
2005 Nonsense mutation in exon 3 of Rpe65 in the rd12 mouse causes absence of RPE65 protein, no 11-cis-retinal, no rhodopsin, and retinyl ester accumulation in the RPE; the rd12 mutation maps to mouse chromosome 3 near the Rpe65 locus. Genetic linkage analysis; direct sequencing; immunohistochemistry; ERG; biochemical retinoid analysis Molecular vision High 15765048
2009 Eight RPE65 missense variants predicted to be disease-causing by an EPP algorithm all showed isomerase activity <6% of wild-type in a cell-culture assay; three variants predicted non-pathogenic showed activities of 68%, 127%, and 110% of wild-type; complete concordance between EPP prediction and functional assay established that most RPE65 disease mutations cause loss of isomerase function. Cell-based isomerase activity assay; site-directed mutagenesis of 11 RPE65 variants; EPP algorithm Human mutation Medium 19431183
2014 The rd12 allele does not complement the Rpe65 knockout allele (complementation test), confirming the rd12 lesion is in Rpe65; the rd12 mutant mRNA is correctly spliced and exported to cytoplasm but is enriched on ribosome-free mRNPs rather than actively translating polyribosomes, indicating translational suppression as the mechanism of absent protein; the rd12 allele exerts a semidominant negative effect on visual function. Complementation cross; sucrose gradient polysome fractionation; RNA FISH; qRT-PCR; ERG; optokinetic tracking; immunoblot Investigative ophthalmology & visual science Medium 24644049
2005 In zebrafish (cone-dominated retina), RPE65 knockdown reduces 11-cis-retinal levels and alters rod outer segment morphology, but cone vision remains functional; all-trans retinylamine (rod visual cycle inhibitor) does not have additive effects in RPE65-deficient larvae, providing in vivo evidence for an RPE65-independent pathway for cone 11-cis-retinal regeneration. Morpholino-mediated gene knockdown in zebrafish; behavioral vision testing; HPLC retinoid profiling; retinylamine pharmacological inhibition The European journal of neuroscience Medium 17868371
1998 A 4-nucleotide (AAGA) deletion in canine RPE65 (nucleotides 487-490) causing a frameshift and premature stop codon is homozygous in RPE65-mutant Briard dogs with congenital stationary night blindness and retinal dystrophy; the mutation causes retinal dysfunction and RPE lipid vacuole accumulation, demonstrating that RPE65 loss of function disrupts the visual cycle and retinal architecture. RT-PCR and cDNA cloning; direct sequencing; ERG; fundus and morphological examination; haplotype analysis Molecular vision Medium 9808841
2018 Cone-expressed RPE65 in mouse cones does not detectably contribute to cone function or morphology in transgenic mice expressing human RPE65 specifically in cones on an RPE65-deficient background; only a slight delay in dark adaptation was observed, possibly due to retinoid buffering. Transgenic mice (cone-specific human RPE65 expression in RPE65-deficient background); ERG dark adaptation; single-cell electrophysiology; morphological analysis Scientific reports Medium 30242264

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2017 Efficacy and safety of voretigene neparvovec (AAV2-hRPE65v2) in patients with RPE65-mediated inherited retinal dystrophy: a randomised, controlled, open-label, phase 3 trial. Lancet (London, England) 1517 28712537
1998 Rpe65 is necessary for production of 11-cis-vitamin A in the retinal visual cycle. Nature genetics 814 9843205
2011 Gene therapy for leber congenital amaurosis caused by RPE65 mutations: safety and efficacy in 15 children and adults followed up to 3 years. Archives of ophthalmology (Chicago, Ill. : 1960) 526 21911650
2005 RPE65 is the isomerohydrolase in the retinoid visual cycle. Proceedings of the National Academy of Sciences of the United States of America 455 16116091
1998 Mutations in the RPE65 gene in patients with autosomal recessive retinitis pigmentosa or leber congenital amaurosis. Proceedings of the National Academy of Sciences of the United States of America 386 9501220
2005 Rpe65 is the retinoid isomerase in bovine retinal pigment epithelium. Cell 369 16096063
2019 Efficacy, Safety, and Durability of Voretigene Neparvovec-rzyl in RPE65 Mutation-Associated Inherited Retinal Dystrophy: Results of Phase 1 and 3 Trials. Ophthalmology 322 31443789
1993 Molecular cloning and expression of RPE65, a novel retinal pigment epithelium-specific microsomal protein that is post-transcriptionally regulated in vitro. The Journal of biological chemistry 265 8340400
2009 Human RPE65 gene therapy for Leber congenital amaurosis: persistence of early visual improvements and safety at 1 year. Human gene therapy 263 19583479
2000 Genetics and phenotypes of RPE65 mutations in inherited retinal degeneration. Investigative ophthalmology & visual science 226 11095629
2010 Leber congenital amaurosis due to RPE65 mutations and its treatment with gene therapy. Progress in retinal and eye research 207 20399883
2003 Functional and structural recovery of the retina after gene therapy in the RPE65 null mutation dog. Investigative ophthalmology & visual science 200 12657607
2005 Retinal degeneration 12 (rd12): a new, spontaneously arising mouse model for human Leber congenital amaurosis (LCA). Molecular vision 193 15765048
2005 Gene therapy restores vision-dependent behavior as well as retinal structure and function in a mouse model of RPE65 Leber congenital amaurosis. Molecular therapy : the journal of the American Society of Gene Therapy 171 16223604
1998 Congenital stationary night blindness in the dog: common mutation in the RPE65 gene indicates founder effect. Molecular vision 154 9808841
2020 Clinical Perspective: Treating RPE65-Associated Retinal Dystrophy. Molecular therapy : the journal of the American Society of Gene Therapy 139 33278565
2007 Human cone photoreceptor dependence on RPE65 isomerase. Proceedings of the National Academy of Sciences of the United States of America 138 17848510
2011 A dominant mutation in RPE65 identified by whole-exome sequencing causes retinitis pigmentosa with choroidal involvement. European journal of human genetics : EJHG 132 21654732
2009 RPE65: role in the visual cycle, human retinal disease, and gene therapy. Ophthalmic genetics 126 19373675
2015 The Status of RPE65 Gene Therapy Trials: Safety and Efficacy. Cold Spring Harbor perspectives in medicine 123 25635059
2017 Safety and Long-Term Efficacy of AAV4 Gene Therapy in Patients with RPE65 Leber Congenital Amaurosis. Molecular therapy : the journal of the American Society of Gene Therapy 110 29033008
2006 Lentiviral gene transfer of RPE65 rescues survival and function of cones in a mouse model of Leber congenital amaurosis. PLoS medicine 102 17032058
2002 Identification of the RPE65 protein in mammalian cone photoreceptors. Investigative ophthalmology & visual science 98 11980880
2007 Canine and human visual cortex intact and responsive despite early retinal blindness from RPE65 mutation. PLoS medicine 97 17594175
2018 Gene therapy for RPE65-related retinal disease. Ophthalmic genetics 92 30335549
2003 In vivo gene therapy in young and adult RPE65-/- dogs produces long-term visual improvement. The Journal of heredity 92 12692160
2019 CRISPR-Cas9-mediated therapeutic editing of Rpe65 ameliorates the disease phenotypes in a mouse model of Leber congenital amaurosis. Science advances 89 31692906
2008 A comprehensive clinical and biochemical functional study of a novel RPE65 hypomorphic mutation. Investigative ophthalmology & visual science 75 18599565
2021 Retinal pigment epithelium 65 kDa protein (RPE65): An update. Progress in retinal and eye research 74 34607013
2006 RPE65 gene delivery restores isomerohydrolase activity and prevents early cone loss in Rpe65-/- mice. Investigative ophthalmology & visual science 73 16505056
2019 Long-Term Structural Outcomes of Late-Stage RPE65 Gene Therapy. Molecular therapy : the journal of the American Society of Gene Therapy 63 31604676
2010 Self-complementary AAV-mediated gene therapy restores cone function and prevents cone degeneration in two models of Rpe65 deficiency. Gene therapy 62 20237510
2010 Gene therapy in the second eye of RPE65-deficient dogs improves retinal function. Gene therapy 61 20703309
2007 RDH12 and RPE65, visual cycle genes causing leber congenital amaurosis, differ in disease expression. Investigative ophthalmology & visual science 61 17197551
2007 RPE65 is essential for the function of cone photoreceptors in NRL-deficient mice. Investigative ophthalmology & visual science 60 17251447
2010 Importance of membrane structural integrity for RPE65 retinoid isomerization activity. The Journal of biological chemistry 59 20100834
2005 Mole quantity of RPE65 and its productivity in the generation of 11-cis-retinal from retinyl esters in the living mouse eye. Biochemistry 59 16026160
2001 Expression, purification, and MALDI analysis of RPE65. Investigative ophthalmology & visual science 58 11381042
2011 Gene therapy rescues cone structure and function in the 3-month-old rd12 mouse: a model for midcourse RPE65 leber congenital amaurosis. Investigative ophthalmology & visual science 57 21169527
2020 The effect of human gene therapy for RPE65-associated Leber's congenital amaurosis on visual function: a systematic review and meta-analysis. Orphanet journal of rare diseases 55 32059734
2012 RPE65 gene therapy slows cone loss in Rpe65-deficient dogs. Gene therapy 54 22951453
2010 Membrane-binding and enzymatic properties of RPE65. Progress in retinal and eye research 53 20304090
2006 Rpe65 as a modifier gene for inherited retinal degeneration. The European journal of neuroscience 51 16519667
2018 Results at 5 Years After Gene Therapy for RPE65-Deficient Retinal Dystrophy. Human gene therapy 48 29869534
2007 Evidence for RPE65-independent vision in the cone-dominated zebrafish retina. The European journal of neuroscience 46 17868371
2002 Clinical course and visual function in a family with mutations in the RPE65 gene. Archives of ophthalmology (Chicago, Ill. : 1960) 44 11786058
2002 Retinal degeneration and RPE transplantation in Rpe65(-/-) mice. Investigative ophthalmology & visual science 44 12356839
2020 Voretigene neparvovec-rzyl for treatment of RPE65-mediated inherited retinal diseases: a model for ocular gene therapy development. Expert opinion on biological therapy 43 32149547
2009 Predicting the pathogenicity of RPE65 mutations. Human mutation 42 19431183
2006 Visual cycle protein RPE65 persists in new retinal cells during retinal regeneration of adult newt. The Journal of comparative neurology 42 16485283
1998 Promoter analysis of RPE65, the gene encoding a 61-kDa retinal pigment epithelium-specific protein. Investigative ophthalmology & visual science 42 9501877
2014 Rescue of enzymatic function for disease-associated RPE65 proteins containing various missense mutations in non-active sites. The Journal of biological chemistry 41 24849605
2009 Evaluation of 9-cis-retinyl acetate therapy in Rpe65-/- mice. Investigative ophthalmology & visual science 41 19407008
2002 Prenatal human ocular degeneration occurs in Leber's congenital amaurosis (LCA2). The journal of gene medicine 40 12124981
1998 Cloning and localization of RPE65 mRNA in salamander cone photoreceptor cells1. Biochimica et biophysica acta 40 9838153
2011 RPE65 is present in human green/red cones and promotes photopigment regeneration in an in vitro cone cell model. The Journal of neuroscience : the official journal of the Society for Neuroscience 39 22171060
2023 Single Center Experience with Voretigene Neparvovec Gene Augmentation Therapy in RPE65 Mutation-Associated Inherited Retinal Degeneration in a Clinical Setting. Ophthalmology 38 37704110
2022 Epidemiology of Mutations in the 65-kDa Retinal Pigment Epithelium (RPE65) Gene-Mediated Inherited Retinal Dystrophies: A Systematic Literature Review. Advances in therapy 38 35098484
1997 RPE65, the major retinal pigment epithelium microsomal membrane protein, associates with phospholipid liposomes. Archives of biochemistry and biophysics 38 9328280
2013 Persistence of non-viral vector mediated RPE65 expression: case for viability as a gene transfer therapy for RPE-based diseases. Journal of controlled release : official journal of the Controlled Release Society 37 24035979
2018 Insights into the pathogenesis of dominant retinitis pigmentosa associated with a D477G mutation in RPE65. Human molecular genetics 35 29659842
2014 Pseudo-fovea formation after gene therapy for RPE65-LCA. Investigative ophthalmology & visual science 34 25537204
2013 Fatty acid transport protein 4 (FATP4) prevents light-induced degeneration of cone and rod photoreceptors by inhibiting RPE65 isomerase. The Journal of neuroscience : the official journal of the Society for Neuroscience 34 23407971
2008 Impact of retinal disease-associated RPE65 mutations on retinoid isomerization. Biochemistry 34 18722466
2006 Two point mutations of RPE65 from patients with retinal dystrophies decrease the stability of RPE65 protein and abolish its isomerohydrolase activity. The Journal of biological chemistry 34 16754667
2019 Generation and Characterization of Induced Pluripotent Stem Cells and Retinal Organoids From a Leber's Congenital Amaurosis Patient With Novel RPE65 Mutations. Frontiers in molecular neuroscience 33 31572124
2013 Successful gene therapy in older Rpe65-deficient dogs following subretinal injection of an adeno-associated vector expressing RPE65. Human gene therapy 33 24028205
2018 Two-year progression analysis of RPE65 autosomal dominant retinitis pigmentosa. Ophthalmic genetics 32 29947567
2021 RPE65-associated inherited retinal diseases: consensus recommendations for eligibility to gene therapy. Orphanet journal of rare diseases 31 34088339
2021 Inherited Retinal Diseases Due to RPE65 Variants: From Genetic Diagnostic Management to Therapy. International journal of molecular sciences 31 34281261
2007 Electroretinographic analyses of Rpe65-mutant rd12 mice: developing an in vivo bioassay for human gene therapy trials of Leber congenital amaurosis. Molecular vision 30 17960108
2023 Classification and Growth Rate of Chorioretinal Atrophy after Voretigene Neparvovec-Rzyl for RPE65-Mediated Retinal Degeneration. Ophthalmology. Retina 29 37660736
2016 A Dominant Mutation in Rpe65, D477G, Delays Dark Adaptation and Disturbs the Visual Cycle in the Mutant Knock-In Mice. The American journal of pathology 28 28041994
2005 RPE65 surface epitopes, protein interactions, and expression in rod- and cone-dominant species. Molecular vision 28 16379027
1999 Identification of RPE65 in transformed kidney cells. FEBS letters 28 10386590
2020 Voretigene Neparvovec: A Review in RPE65 Mutation-Associated Inherited Retinal Dystrophy. Molecular diagnosis & therapy 27 32535767
2012 Novel RPE65 mutations associated with Leber congenital amaurosis in Chinese patients. Molecular vision 26 22509104
2019 Aberrant RNA splicing is the major pathogenic effect in a knock-in mouse model of the dominantly inherited c.1430A>G human RPE65 mutation. Human mutation 25 30628748
2003 Gene transfer in the RPE65 null mutation dog: relationship between construct volume, visual behavior and electroretinographic (ERG) results. Documenta ophthalmologica. Advances in ophthalmology 25 12906125
2022 Human primitive mesenchymal stem cell-derived retinal progenitor cells improved neuroprotection, neurogenesis, and vision in rd12 mouse model of retinitis pigmentosa. Stem cell research & therapy 22 35395806
2002 Light damage susceptibility and RPE65 in rats. Experimental eye research 22 12387788
2000 A homozygous deletion in RPE65 in a small Sardinian family with autosomal recessive retinal dystrophy. Molecular vision 22 11062306
2020 Cost Effectiveness of Voretigene Neparvovec for RPE65-Mediated Inherited Retinal Degeneration in Germany. Translational vision science & technology 21 32879773
2019 RPE65 mutation frequency and phenotypic variation according to exome sequencing in a tertiary centre for genetic eye diseases in China. Acta ophthalmologica 21 31273949
2005 Function of the protein RPE65 in the visual cycle. Nutrition reviews 21 15825812
2021 RPE65-related retinal dystrophy: Mutational and phenotypic spectrum in 45 affected patients. Experimental eye research 20 34492281
2019 The dual roles of RPE65 S-palmitoylation in membrane association and visual cycle function. Scientific reports 20 30914787
2018 Examining the Role of Cone-expressed RPE65 in Mouse Cone Function. Scientific reports 20 30242264
2004 RPE65 of retinal pigment epithelium, a putative receptor molecule for plasma retinol-binding protein, is expressed in human keratinocytes. The Journal of investigative dermatology 20 15009723
2022 RPE65-Associated Retinopathies in the Italian Population: A Longitudinal Natural History Study. Investigative ophthalmology & visual science 19 35129589
2018 A novel RPE65 inhibitor CU239 suppresses visual cycle and prevents retinal degeneration. Biochimica et biophysica acta. Molecular basis of disease 19 29684583
2010 Cleavage of the retinal pigment epithelium-specific protein RPE65 under oxidative stress. International journal of biological macromolecules 19 20510285
2019 Utility of In Vitro Mutagenesis of RPE65 Protein for Verification of Mutational Pathogenicity Before Gene Therapy. JAMA ophthalmology 18 31580392
2019 Pathogenicity Reclasssification of RPE65 Missense Variants Related to Leber Congenital Amaurosis and Early-Onset Retinal Dystrophy. Genes 18 31878136
2017 Evaluation of tolerance to lentiviral LV-RPE65 gene therapy vector after subretinal delivery in non-human primates. Translational research : the journal of laboratory and clinical medicine 18 28754419
2015 Investor Outlook: Significance of the Positive LCA2 Gene Therapy Phase III Results. Human gene therapy. Clinical development 18 26684444
2012 Vitamin A metabolism in benign and malignant melanocytic skin cells: importance of lecithin/retinol acyltransferase and RPE65. Journal of cellular physiology 18 21465477
2011 Clinical gene therapy for the treatment of RPE65-associated Leber congenital amaurosis. Expert opinion on biological therapy 17 21299439
2015 Investor Outlook: Focus on Upcoming LCA2 Gene Therapy Phase III Results. Human gene therapy. Clinical development 16 26390089
2014 The Rpe65 rd12 allele exerts a semidominant negative effect on vision in mice. Investigative ophthalmology & visual science 16 24644049

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